Chromatographic Column For Separating Exosomes （Cell Supernatant）

Exosomes exist naturally in body fluids such as blood, saliva, urine, and breast milk. They are nanoscale membranous vesicles with uniform size and particle size of 30 to 150 nm that are actively secreted by cells into the extracellular space. This vesicle has its own specific markers, and is coated with proteins, mRNA, miRNA, lipids, and other substances, which can be used as a pathway for substance and signal communication between cells and participate in processes of cell growth, cell proliferation, cell differentiation, cell migration, angiogenesis, and tumor growth.

This product is a size exclusion chromatography column (SEC) suitable for rapid separation and purification of serum/plasma exosomes. It can retain high-purity and highly naturally active exosomes while efficiently removing proteins; Effectively avoiding the damage to the extracellular membrane structure caused by ultra-high centrifugal force in traditional ultracentrifugation methods, while reducing the impact of high concentration proteins and other substances on the purity of exosomes in traditional precipitation methods.

Transportation at room temperature; Store at room temperature with a validity period of 12 months.

1. Preparation

a) Prepare a room temperature, sterile buffer solution, such as PBS (recommended G4202);

b) Prepare room temperature, sterile 0.1M NaOH reagent and 20% ethanol;

c) Before using the separation column, remove the red upper cover of the separation column and place it on a bracket (recommended G6063) for use;

2. Exosomes separation

a) Using serum/plasma biological samples 0.22 μm membrane filtration, preliminary purification for backup;

b) Remove the white bottom cover of the separation column, and after the protective liquid inside the column flows out, add a total of 30 mL buffer solution liquid in batches to clean the separation column;

c) After all the buffer above the sieve plate enters the separation column; Take 0.5mL of the backup serum/plasma from step 2.a) and add it to the center of the sieve plate (if less than 0.5mL, add buffer solution to make up to 0.5mL);

d) After all serum/plasma passes through the sieve plate and enters the separation column, add 2.5mL buffer solution;

e) After all the buffer solution in the previous step passes through the sieve plate and enters the separation column, add 1 mL of buffer solution. At the same time, collect 1 mL of washout solution, which is rich in naturally active exosomes and can be used for subsequent experiments or frozen at -80 ℃ for backup;

3. Post separation treatment

a) After completing the separation of exosomes, add a total of 30 mL of buffer solution in batches to clean the separation column;

b) If it is necessary to continue using the separation column to separate serum/plasma exosomes, repeat steps 2.c) to 2.e);

c) If it is not necessary to continue using the separation column to separate serum/plasma exosomes, first add 5 mL of 0.1M NaOH to clean the separation column, then add a total of 20 mL of buffer solution in batches to clean and balance the separation column, and then add 5 mL of 20% ethanol to clean the separation column. After cleaning, cover the white bottom cover of the separation column; Finally, add 2mL of 20% ethanol to moisturize, cover and tighten the red top of the separation column, and store the separation column at room temperature.

1. When separating exosomes from different serum/plasma, the elution curve and purity of this product may slightly differ, but it does not affect the overall trend.

2. When the sample size is large or the content of exosomes in the sample is low, the sample can be concentrated first and then separated using a separation chromatography column.

3. During the process of separating serum/plasma exosomes from this product, it is necessary to avoid prolonged interruption of liquid flow in the separation column.

4. During the use of this product, do not separate biological samples from different sources on the same column, and do not reuse the same column more than 5 times.

5. This product is suitable for storage and use at room temperature, and low temperature use and high temperature heating should be avoided.

6. During the experiment, please wear lab clothes and disposable gloves to avoid contamination and ensure safety.

For Research Use Only!